ABSTRACT HAIRBASE: THE DEVELOPMENT OF AN ONLINE REFERENCE ATLAS OF MAMMALIAN HAIR The class Mammalia contains approximately 5,400 species that display incredible variation in their pelage characteristics. When attempting to identify an unknown animal based on the morphological characteristics of its hair, the macroscopic and microscopic examination of guard hairs is often used. Species identification from small amounts of hair or single hairs has been the goal of many published atlases. However, many of these atlases do not adequately reflect the range of variation both between and within individual species. HAIRbase™ is an online digital database of bright field and scanning electron microscope images of mammalian hair. In contrast to many other atlases, HAIRbase™ contains images from both primary and secondary guard hairs from three distinct regions on the body from each species represented. HAIRbase™ is an applicative tool that can aid forensic hair examiners by providing a visual reference that contains diagnostic information regarding traits of the hair shield, subshield, and basal regions, including medullary configurations, cortical characteristics, and cuticular patterns that can be used to generate leads in investigations involving animals. Michael Vincent Gonzalez May 2011 HAIRBASE: THE DEVELOPMENT OF AN ONLINE REFERENCE ATLAS OF MAMMALIAN HAIR by Michael Vincent Gonzalez A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science in Forensic Science in the College of Science and Mathematics California State University, Fresno May 2011 © 2011 Michael Vincent Gonzalez APPROVED For the Department of Chemistry: We, the undersigned, certify that the thesis of the following student meets the required standards of scholarship, format, and style of the university and the student's graduate degree program for the awarding of the master's degree. Michael Vincent Gonzalez Thesis Author Kevin W.P. Miller (Chair) Chemistry James P. Prince Biology Bonnie Yates National Fish and Wildlife Forensic Laboratory Margaret E. Sims National Fish and Wildlife Forensic Laboratory For the University Graduate Committee: Dean, Division of Graduate Studies AUTHORIZATION FOR REPRODUCTION OF MASTER’S THESIS X I grant permission for the reproduction of this thesis in part or in its entirety without further authorization from me, on the condition that the person or agency requesting reproduction absorbs the cost and provides proper acknowledgment of authorship. Permission to reproduce this thesis in part or in its entirety must be obtained from me. Signature of thesis author: ACKNOWLEDGMENTS This thesis has been a long and arduous journey beset with many difficulties. As it stands complete, I can only say, thank you to my strong support system of family, friends, and co-workers that not only helped make this project the success that it was, but also helped me keep my sanity during those tough days in the laboratory. First, I would like to thank my mother and father, without them, I would not be here (both literally and figuratively). I would like to thank my father for instilling in me the benefit and importance of a strong education. Also, I would like to thank him for always being in support of any crazy dream that seemed to pop into my head (even if it meant moving to Fresno to pursue my Master’s degree in Forensic Science). I would also like to thank my mother, who has been a pillar of support through good times and bad. She has always been the one reminding me to stay humble, even in the face of educational success. I would like to thank her for genuinely believing that I can do anything that I set my mind to, and then convincing me to go and do it! I would also like to thank my twin sister Casey, a large part of my cheering section, and someone who thinks the world of me. Always calling me the “smart twin” (despite her own educational success) and always being a willing ear to listen in any circumstance. She has been my voice of reason through all the trials and tribulations of graduate school (and life for that matter). Honest to a fault, she has always been ready with what I needed (not always what I wanted) to hear. I don’t believe that I could have made it through without her in my corner. I would like to thank and express my deepest gratitude to Ms. Elsbeth Murata – my right hand in the laboratory and also one of my best and closest vi friends. It would be an understatement to say that she merely “helped” me with this project. She took equal part in the development of HAIRbase™, often willing to give more of her time and resources to this project than I was. Without her, this project would not be half of what it is today. I have seen her grow from an inexperienced undergraduate into quite an accomplished hair morphologist in her own right. I know that she will go on to do great things in the sciences. She has been one of my biggest supporters and always knew that I could do it (even when I doubted myself). I want to thank her for not only putting so much tireless effort into this project, but also for putting up with me (way more than she probably wanted to). Hours of imaging of individual hairs, marathon photo-shopping sessions that went into the wee hours of the morning, she was there through it all (and to my surprise, didn’t want to leave!). I know that a mere thanks in an acknowledgement section of a thesis does not encompass the amount of credit that she deserves, but again, thank you so much! I would like to personally thank Maria Carrizales for her tireless efforts to buoy my spirits after long, rough days and being a strong support system that I could lean on in times of trouble. She was always ready at a moment’s notice to edit any writing, listen to a speech, or answer any question that I had (despite her own work dealing with her PhD). She is the definition of a true friend. I would like to thank her for always being there, and also, for giving me “tough love” when I really needed it. I would like to thank my thesis committee for their efforts to keep me on track and staying positive. I would like to thank Bonnie Yates and Cookie Sims for teaching me not only the finer points of hair morphology and trace analysis, but also how my project would function in the professional realm. They always believed that I could finish and that I would go on to do great things! Both of them vii were always in my corner and were a large part of my cheering section, even when deadlines were looming and things looked their bleakest. I would like the thank Dr. Prince for being my “non forensic committee member”. Approaching the project from a strictly biological sense gave the project more depth and made it a more valuable resource. I want to thank him for his efforts as an editor and as a great source for feedback of a different perspective. I would like to thank Dr. Kevin Miller for standing by my side during every iteration and troubleshooting endeavor that came with this project. I want to thank him for always making himself available should the need arise and planting it in my head that I had what it took to get my PhD. He has helped me, not only to succeed professionally, but also in my personal life as well. He has been there through thick and thin (kidney failure being a particularly “thin” time). He has been one of my biggest cheerleaders throughout my graduate school journey. I also wanted to thank him for pushing me to be more than even I thought I could be, he has truly been instrumental in my success. I would like to thank the California State University, Fresno Biology Department and the National Fish and Wildlife Forensic Laboratory for donating hair samples to this project. To all the other people (you know who you are) that were there to help me out and encourage me, I give a very sincere thanks! TABLE OF CONTENTS Page LIST OF FIGURES .................................................................................................. x INTRODUCTION .................................................................................................... 1 LITERATURE REVIEW ......................................................................................... 6 Hair Physiology ................................................................................................. 6 Macroscopic Variation of Hair .......................................................................... 8 Microscopic Variation of Hair ......................................................................... 10 Hair Atlases and Classification Schemes ........................................................ 12 MATERIALS AND METHODS ........................................................................... 17 Sample Collection ........................................................................................... 17 Loose Hair Storage .......................................................................................... 18 Preparation of Hair for Microscopical Examination ....................................... 19 Microscopy Slide and Stub Storage ................................................................ 21 Digital Image Acquisition ............................................................................... 21 Photo Manipulation ......................................................................................... 24 Development of an Internet-based User Interface ........................................... 25 RESULTS AND DISCUSSION ...........................................................................